Fully automated two-dimensional electrophoresis system for high-throughput protein analysis

We developed a fully automated electrophoresis system for rapid and highly reproducible protein analysis. All the two-dimensional (2D) electrophoresis procedures including isoelectric focusing (IEF), on-part protein staining, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and in situ protein detection were automatically completed. The system comprised Peltiert devices, high-voltage generating devices, electrodes, and three disposable polymethylmethacrylate (PMMA) parts for IEF, reaction chambers, and SDS-PAGE. Because of miniaturization of the IEF part, rapid IEF was achieved in 30 min. A gel with a tapered edge gel on the SDS-PAGE part realized a connection between the parts without use of a gluing material. A biaxial conveyer was employed for the part relocation, sample introduction, and washing processes to realize a low-maintenance and cost-effective automation system. Performances of the system and a commercial minigel system were compared in terms of detected number, resolution, and reproducibility of the protein spots. The system achieved high-resolution comparable to the minigel system despite shorter focusing time and smaller part dimensions. The resulting reproducibility was better or comparable to the performance of the minigel system. Complete 2D separation was achieved within 1.5 h. The system is practical, portable, and has automation capabilities.     

Mechanical properties of textile-inserted PP/PP knitted composites using injection–compression molding

This paper concentrates on the experimental investigation of the self-reinforced all-polypropylene composites. There exists an optimum processing condition to produce high quality specimens by injection–compression molding. Tensile and 3-point bending properties of the virgin PP materials were nearly unaffected by the introduction of reinforcing knit layer(s) due to very low fibre content of the knitted fabrics used. 3-point bending properties were also unaffected by the surface of indentation-flexure. The applied impact energy was maintained at 5 J for the homo-PP and 27 J for the block-PP materials, respectively, to cause penetration during drop-weight impact tests. It is interestingly noteworthy that the self-reinforced homo-PP composites exhibited superior energy absorption capability when compared with the virgin matrix materials. The corresponding plate bending performances of the self-reinforced homo-PP composites also revealed consistent improvement as compared to their virgin counterparts. On the other hand, although virgin block-PP material exhibited better impact performances than its composite reinforced by the homo-PP knitted fabric, a notably small increase in the reinforcement fibre content revealed considerable improvement in the impact properties comparable to those of the virgin block-PP matrix materials. These self-reinforced homo-PP/block-PP materials have clearly indicated that they have the potential to out-perform the block-PP materials via modification and/or manipulation of the reinforcement knit structural/geometrical parameters and the content of reinforcement fibres. Both static and dynamic impact properties are likely to be affected by the local area properties of the tested face under indentation, and thereby contributing to the improved performances of the composite specimens with the knit face under the impact.     

Preparation of plastic spherical microlenses by use of a fluoropolymer stencil and oil-bath heating

A new method for fabricating plastic spherical microlenses was developed, which allowed self-alignment of lenses and self-organized formation of a spherical shape. First a low-surface-energy fluoropolymer thin film was deposited and patterned as a stencil. Then photosensitive phenol resin was patterned on it as the lens material. Finally the resin was annealed in an oil bath to form a sphere. The molten phenol resin spontaneously formed a sphere and positioned itself in the center of the fluoropolymer ring pattern as a result of the difference of surface free energy and the equivalently zero-gravity condition in the oil bath. When a light-emitting-diode printer head was loaded with spherical microlenses, its optical output increased by 1 order of magnitude.     

A CMOS integrated circuit for multichannel multiple-subjectbiotelemetry using bidirectional optical transmissions [weightlessnessin space effects application]

A CMOS integrated circuit for a noninvasive biological-signal telemetry system specified for use in medical and physiological studies of the influence of weightlessness in space is presented. The system can monitor multichannel (4 channels maximum) biological signals from multiple subjects (4 subjects maximum) in real time by using time multiplexing. A key technique so-called synchronized multiple-subject telemetry, to achieve multiple-subject telemetry has been proposed. This technique utilizes bidirectional optical transmissions with direct and scattered infrared lights between an observer and each of the subjects. An experimental CMOS IC to give a small light-weight low-power, and smart telemetry instrument for use on animals has been developed. This IC is for evaluating circuit blocks of the implantable monolithic telemetry instrument. The major circuit blocks include CMOS digital circuits for synchronization, subject selection and time multiplexing, analog circuits for pulse interval modulation, and other blocks such as a CMOS optical pulse receiver and an LED driver. A preliminary experimental multichannel telemetry from two subjects has been performed with the implemented IC chips, and the principal operation of the multiple-subject optical biotelemetry has been demonstrated     

Serotonin 5-HT4 receptor agonistic activity of the optical isomers of (+/-)-4-amino-N-[2-(1-azabicyclo[3.3.0]octan-5-yl)ethyl]-5-chloro-2,3- dihydro-2-methylbenzo[b]furan-7-carboxamide

OBJECTIVE: To compare the influence of incongruent (asymmetric) follicular development on treatment outcome in IVF-ET and GIFT cycles. DESIGN: A retrospective comparative study. SETTING: Tertiary referral center for infertility. PATIENT(S): Five hundred forty-three consecutive assisted reproduction cycles (428 IVF-ET and 115 GIFT) in 422 infertile patients. INTERVENTION(S): Controlled ovarian hyperstimulation (COH) and IVF-ET or GIFT. MAIN OUTCOME MEASURE(S): The incongruity ratio as a parameter of the asymmetry in follicular development and pregnancy rate (PR). RESULT(S): For GIFT cycles, the PRs were 37.8% and 15.7% in cycles with congruent and incongruent follicular development, respectively. However, for IVF-ET cycles, the PR was not affected by incongruent follicular development: 28.2% and 29.0%, respectively. An inverse relationship was observed between the degree of incongruity and the estimated probability of pregnancy in GIFT cycles but not in IVF-ET cycles. Neither the side of the dominant ovary nor the degree of incongruity were consistent in consecutive cycles. CONCLUSION(S): Incongruent follicular development during COH has a significantly negative influence on the outcome of GIFT cycles but not on the outcome of IVF-ET cycles. The reason for this difference is not clear. We recommend considering IVF-ET instead of GIFT if incongruent follicular development occurs.     

Desensitization of the GABAA receptor shifts the dynamic range of retinal horizontal cells due to light and dark adaptation

The effects of the GABAergic pathway on the dynamic range of the light response of cone-driven horizontal cells (HCs) in the carp retina were investigated using conventional intracellular recordings and computer simulations. The amplitude of the HC responses to repetitive test flashes was altered by background illumination in control Ringer's solution, but was not changed following the application of picrotoxin or bicuculline, which suggests that GABAA receptors mediate the subsequent change in light response. This phenomenon was simulated by introducing the desensitization mechanism of the GABAA receptor into a cone-HC network model. These results suggest that desensitization of the GABAA receptor shifts the dynamic range of the HCs due to light and dark adaptation.     

Determination of standard gibbs energies of formation of Cr2N and CrN

The standard Gibbs energies of formation of Cr₂N and CrN have been measured by an equilibration technique and by using thermogravimetry and differential thermal analysis (TG-DTA) at temperatures ranging from 1232 to 1523 K. The results are expressed as follows:

Three-dimensional organization of collagen fibrils during corneal stromal wound healing after excimer laser keratectomy

PURPOSE: To investigate the structural changes in corneal stromal collagen fibrils after excimer laser keratectomy in relation to the degree of corneal haze. SETTING: University of Tokyo Hospital, Tokyo, Japan. METHODS: Corneal haze was quantitatively measured by analyzing the light scattering in Scheimpflug images of the corneas of white rabbits after excimer laser keratectomy. Collagen fibril structure was examined using scanning electron microscopy after chemical digestion with sodium hydroxide solution; the same specimens were examined by transmission electron microscopy after re-embedding. RESULTS: Corneal haze reached a peak 4 weeks after excimer laser keratectomy and then gradually decreased. The collagen fibrils of the normal cornea were regularly arranged parallel to the surface of the cornea, with small interfibrillar distances. After excimer laser keratectomy, the arrangement was highly disordered, with increased interfibrillar distances. These structural changes were most prominent 4 weeks after excimer laser keratectomy. CONCLUSION: The structural changes in the collagen fibrils of the corneal stroma, especially the increase in interfibrillar distances and the disordered arrangement, were associated with corneal haze after excimer laser keratectomy.     

Involvement of heat shock protein 90 in the degradation of mutant insulin receptors by the proteasome

We previously reported three families with type A insulin-resistant syndrome who had mutations, either Asp1179 or Leu1193, in the kinase domain of the insulin receptor. The extreme insulin resistance of these patients was found to be caused by the decreased number of insulin receptors on the cell surface, due to the intracellular rapid degradation (Imamura, T., Takata, Y., Sasaoka, T., Takada, Y., Morioka, H., Haruta, T., Sawa, T., Iwanishi, M., Yang, G. H., Suzuki, Y., Hamada, J., and Kobayashi, M. (1994) J. Biol. Chem. 269, 31019-31027). In the present study, we first examined whether these mutations caused rapid degradation of unprocessed proreceptors, using the exon 13 deleted mutant insulin receptors (DeltaEx13-IR), which were accumulated in the endoplasmic reticulum as unprocessed proreceptors. The addition of Asp1179 or Leu1193 mutation to DeltaEx13-IR caused accelerated degradation of the unprocessed DeltaEx13-IR in the transfected COS-7 cells. Next, we tested whether these mutant receptors were degraded by the proteasome. Treatment with proteasome inhibitors Z-Leu-Leu-Nva-H (MG-115) or Z-Leu-Leu-Leu-H (MG-132) prevented the accelerated degradation of these mutant receptors, resulting in increased amounts of the mutant receptors in the COS-7 cells. Essentially the same results were obtained in the patient's transformed lymphocytes. Finally, we found that these mutant receptors bound to heat shock protein 90 (Hsp90). To determine whether Hsp90 played an important role in the accelerated receptor degradation, we examined the effect of anti-Hsp90 antibody on the mutant receptor degradation. The microinjection of anti-Hsp90 antibody into cells prevented the accelerated degradation of both Asp1179 and Leu1193 mutant insulin receptors. Taken together, these results suggest that Hsp90 is involved in dislocation of the mutant insulin receptors out of the endoplasmic reticulum into the cytosol, where the mutant receptors are degraded by the proteasome.     

Validity of sleep log compared with actigraphic sleep-wake state

OBJECTIVE: To evaluate the efficacy and complications of external cephalic version in term (37 or more weeks) gestation. STUDY DESIGN: A case series from February 1990 until December 1994 studied 160 patients with term singleton breech presentation. External cephalic version (ECV) with prophylactic tocolysis was the method used. RESULTS: ECV was successful in 67% of the cases (107/160) overall, in 56% of the nulliparous (57/102) and 86% of the multiparous women (50/58). About 90% of those with successful ECV (96/107) had a vaginal delivery with a vertex presentation of the fetus. Emergency Caesarean section due to fetal distress was not required during or immediately after the procedure. No perinatal mortality or babies with an Apgar score less than 7 at 5 min were observed. CONCLUSIONS: ECV in term pregnancy seems to be useful and it is safe both for the mother and the fetus. It should be performed at term, with close monitoring of the fetus, and in an environment able to provide an emergency Caesarean section.